• 제목/요약/키워드: confocal microscopy

검색결과 437건 처리시간 0.031초

Analysis of C. elegans VIG-1 Expression

  • Shin, Kyoung-Hwa;Choi, Boram;Park, Yang-Seo;Cho, Nam Jeong
    • Molecules and Cells
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    • 제26권6호
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    • pp.554-557
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    • 2008
  • Double-stranded RNA (dsRNA) induces gene silencing in a sequence-specific manner by a process known as RNA interference (RNAi). The RNA-induced silencing complex (RISC) is a multi-subunit ribonucleoprotein complex that plays a key role in RNAi. VIG (Vasa intronic gene) has been identified as a component of Drosophila RISC; however, the role VIG plays in regulating RNAi is poorly understood. Here, we examined the spatial and temporal expression patterns of VIG-1, the C. elegans ortholog of Drosophila VIG, using a vig-1::gfp fusion construct. This construct contains the 908-bp region immediately upstream of vig-1 gene translation initiation site. Analysis by confocal microscopy demonstrated GFP-VIG-1 expression in a number of tissues including the pharynx, body wall muscle, hypodermis, intestine, reproductive system, and nervous system at the larval and adult stages. Furthermore, western blot analysis showed that VIG-1 is present in each developmental stage examined. To investigate regulatory sequences for vig-1 gene expression, we generated constructs containing deletions in the upstream region. It was determined that the GFP expression pattern of a deletion construct (${\Delta}-908$ to -597) was generally similar to that of the non-deletion construct. In contrast, removal of a larger segment (${\Delta}-908$ to -191) resulted in the loss of GFP expression in most cell types. Collectively, these results indicate that the 406-bp upstream region (-596 to -191) contains essential regulatory sequences required for VIG-1 expression.

Characteristics of chain-forming cells in Gyrodinium impudicum using fluorescent ConA

  • Park, Eun-Ju;Kim, Gwang-Hoon;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제34권4호
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    • pp.214-219
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    • 1999
  • Cells of the dinoflagellate Gyrodinium impudicum form characteristic chains, which are associated with sugar accumulated on the cell surface. To resolve the relationship between chain-formation and cell surface sugar accumulation, confocal microscopy was used to observe sugar accumulation points in the vegetative cells and long chain-forming cells of G. impudicum cells treated with fluorescent-tagged ConA. In the stationary and exponential phases, both vegetative cells and chain-forming cells were similar to each other in fluorescent intensity. There was no evidence that chain-forming cells had a specific location for sugar accumulation on the cell surface. Most of the cells formed 2-cell chains one day after inoculation, but longer chains consisting of 4-8 cells increased markedly in 4day and 8 day cultures. Gyrodinium impudicum chains usually consist of more than four cells, and chains of 8 or even 16 cells can be observed in mature cultures. Temperature played an importantrole in chain-formation of the cells, threshold temperature for the development of long chain-formation was 19 $^{\circ}$C.

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Vasodilation of BCT is Associated with Inhibition of PKC$\alpha$ Translocation and LC20 Phosphorylation

  • Kwon Oh Kui;Shin Dong Hoon;Kim Gil Whon;Shin Heung Mook
    • 동의생리병리학회지
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    • 제17권5호
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    • pp.1335-1338
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    • 2003
  • We have previously reported that the vasodilatory effect of BanhabackchuIChunma-tang(半夏白朮天麻湯; BCT), a herbal formula, and its mechanism might be associated with at least in part NO pathway. In the present study, we studied the influence of BanhabackchulChenuma-tang (BCT) on the phosphorylation of LC20, in parallel, the distribution of α-protein kinase C(PKCα) by phenylephrine was monitored using laser scan confocal immunofluorescent microscopy in freshly isolated ferret portal vein smooth muscle living single cells. Phenylephrine stimulation induced LC20 phosphorylation and translocation of PKCα. However, BCT dephosphorylated LC20 phosphorylation and inhibited the translocation of PKCα. Our results demonstrate that the mechanism of relaxant effect of BanhabackchulChunma-tang inhibition is associated with inhibition of PKCα activation and LC20 phosphorylation.

Platelets Induce Proliferation of Human Umbilical Vein Endothelial Cells via CD154-CD40 Pathway Independently of VEGF

  • Cho, Wha-Jung;Ko, Eun-Mi;Cheon, In-Su;Jeoung, Doo-Il;Kim, Young-Myeong;Choe, Jong-Seon
    • IMMUNE NETWORK
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    • 제8권3호
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    • pp.75-81
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    • 2008
  • Background: Platelets take part in repairing the lesions of endothelial damage. To understand the molecular mechanism of this process, we tested the hypothesis that CD154 expressed on activated platelets stimulates proliferation of human endothelial cells. Methods: The expression levels of CD154 and CD40 on platelets and endothelial cells, respectively, were measured by flow cytometry and confocal microscopy. Function-blocking monoclonal antibody against CD154 was developed after immunization with CD154-transfected L cells. Results: An anti-CD40 agonist antibody and soluble CD154 both induced significant proliferation of endothelial cells. In addition, a function-blocking anti-CD154 antibody inhibited the platelet-induced proliferation of endothelial cells, indicating that the CD154-CD40 pathway is involved in these cellular interactions. An anti-VEGF antibody failed to inhibit the proliferation. This, in addition to the fact that very small amounts of VEGF are released from platelets or endothelial cells, suggests that VEGF does not play an important role in the platelet-stimulated proliferation of endothelial cells. Conclusion: Our results indicate that platelets induce proliferation of endothelial cells by CD154-CD40 interactions independently of VEGF.

Collagen과 osteopontin이 도입된 고기능성 PCL membrane의 제조 및 지방유래 줄기세포의 골세포 분화능 관찰

  • 김영희;송호연;;변인선;박민주;이병택
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2010년도 춘계학술발표대회
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    • pp.42.1-42.1
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    • 2010
  • Poly-carprolactone (PCL)은 생분해성 고분자로 장기간의 임상실험 결과 생체에 독성이 없으며 생체친화성이 우수한 소재로 확인되어 PLGA, PLLA 등과 더불어 조직공학 분야에서 널리 사용되고 있는 생체재료이다. 그러나 PCL은 5개의 비극성 methylene group과 1개의 극성 ester group이 반복되는 지방족의 polyester로 구조상 탄소수가 많아 소수성을 띄는 단점을 가지고 있다. 이러한 표면이 소수성인 재료의 경우, 초기 단백질 흡착능이 떨어져 세포의 부착이 느린 속도로 일어나므로 세포 분화 및 조직 재생이 더디게 일어난다. 본 연구에서는 소수성의 PCL 표면의 단백질 흡착능을 증가시키기 위해 기능성 amine group을 부착하였으며, 또한 골재생을 촉진시킬 수 있는 세포외 기질인 collagen과 osteopontin을 부착함으로써 고기능성 PCL membrane을 제조하였다. 제조된 PCL membrane은 골재생용 조직공학에의 응용을 위해 지방유래 줄기세포를 이용하여 부착능 및 골세포로의 분화능을 확인하였다. 표면 성질의 변화에 의한 세포의 부착능의 변화를 confocal microscopy을 이용하여 부착에 관여하는 단백질의 발현을 확인하였으며, collagen과 osteopontin에 의한 골세포로의 분화능을 확인하기 위해 real time PCR을 통해 골세포의 분화 표지 유전자의 발현을 비교 분석하였다.

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Surface Modification of Magnetites Using Maltotrionic Acid and Folic Acid for Molecular Imaging

  • Selim, K.M.Kamruzzaman;Lee, Joo-Hee;Kim, Sun-Jung;Xing, Zhicai;Kang, Inn-Kyu;Chang, Yong-Min;Guo, Haiqing
    • Macromolecular Research
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    • 제14권6호
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    • pp.646-653
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    • 2006
  • Highly hydrophilic, uniform, superparamagnetic and nontoxic maltotrionic acid (MA)-coated magnetite nano-particles (MAM) were prepared and characterized by TEM, DLS, XRD and VSM. MA was used to improve the biocompatibility, monodispersity and non-specific intracellular uptake of nanoparticles. Folic acid (FA) was subsequently conjugated to the MAM to preferentially target KB cells (cancer cells) that have folate receptors expressed on their surfaces and to facilitate nanoparticles in their transit across the cell membrane. Finally, fluorescence isothiocyanate (FITC) was added to the nanoparticles to visualize the nanoparticle internalization into KB cells. After the cells were cultured in a media containing the MAM and MAM-folate conjugate (FAMAM), the results of fluorescence and confocal microscopy showed that both types of nanoparticles were internalized into the cells. Nevertheless, the amount of FAMAM uptake was higher than that of MAM. This result indicated that nanoparticles modified with MA and FA could be used to facilitate the nanoparticle uptake to specific KB cells (cancer cells) for molecular imaging.

Specific Detection of DNA Using Quantum Dots and Magnetic Beads for Large Volume Samples

  • Kim, Yeon-Seok;Kim, Byoung-Chan;Lee, Jin-Hyung;Kim, Jung-Bae;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권5호
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    • pp.449-454
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    • 2006
  • Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples. In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals. The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs. In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated. The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis. It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal. In addition, the presence of low copies of target DNAs such as 0.5 pM in large volume samples up to 40mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold.

벼 잎 표면에서 액적의 이방성 흐름 특성에 관한 연구 (A Study on the Anisotropic Flow Characteristics of Droplets on Rice Leaf Surface)

  • 김태완
    • Tribology and Lubricants
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    • 제33권6호
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    • pp.251-255
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    • 2017
  • In this study, we aimed to clarify the wettability and anisotropic flow characteristics of rice leaves as a basic study for engineering applications of anisotropic flow characteristics of rice leaf surface. To investigate the surface structure of rice leaf, the micro grooves and asperities of rice leaves were analyzed and quantified by scanning electron microscope, Confocal laser scanning microscopy, and stylus profilometer. The analysis of the structure of rice leaf surface confirmed that asymmetrical cone - like protrusions in leaf veins were inclined toward the leaf tip. The static contact angle test showed that the contact angle at the midline vein or leaf vein location where the micropapilla is concentrated is about $20^{\circ}$ higher than the leaf blade position. The contact angles of fresh and dried rice leave were also compared. The dried rice leaves showed a contact angle of about $5^{\circ}$ to $15^{\circ}$ higher than that of fresh leaves, suggesting that the volume of the protrusions decreased as the water was removed, thus reducing the contact area with the droplet. In the contact angle history test the hysteresis in the leaf tip direction was found to be much lower than that in the leaf petiole direction. This results can be explained that asymmetrical cone - like protrusions had a significant effect on the droplet flow characteristics through contact angle hysteresis experiment.

Use of Cationic PAM as a Surface Sizing Additive to Improve Paper Properties

  • Seo, Man-Seok;Lee, Hak-Lae;Youn, Hye-Jung
    • 한국펄프종이공학회:학술대회논문집
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    • 한국펄프종이공학회 2006년도 PAN PACIFIC CONFERENCE vol.2
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    • pp.245-250
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    • 2006
  • This study was focused on the use of cationic PAM (Polyacrylamide) as a surface sizing additive to improve the surface sizing properties of paper. Effects of the ionic property, viscosity and charge density of PAM on bending stiffness of surface sized papers were investigated. Use of cationic PAM as a surface sizing additive improved bending stiffness while addition of anionic PAM did not show any effect. Increase of starch holdout with the addition of cationic PAM was attributed as a prime reason of stiffness increase. Viscosity of PAM was one of the most important factors affecting surface sizing due to its influence on the interaction between cationic PAM and oxidized starch solution. Greater improvement of bending stiffness of paper was obtained when high charged PAM was used as an additive. The order of addition was found to have significant influence on the effect of additives since it influences the formation of network structure among starch, cationic PAM, and SA (styrene acrylic acid copolymer). Investigation on the penetration of starch solution was carried out with CLSM (Confocal Laser Scanning Microscopy), and it was shown that the addition of cationic PAM to oxidized starch solution made starch molecules stay on the paper surface rather than penetrating into the paper structure because of the electrostatic interaction between negatively charged fibers and positively charged cationic PAM.

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Plastid Transformation in the Monocotyledonous Cereal Crop, Rice (Oryza sativa) and Transmission of Transgenes to Their Progeny

  • Lee, Sa Mi;Kang, Kyungsu;Chung, Hyunsup;Yoo, Soon Hee;Ming Xu, Xiang;Lee, Seung-Bum;Cheong, Jong-Joo;Daniell, Henry;Kim, Minkyun
    • Molecules and Cells
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    • 제21권3호
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    • pp.401-410
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    • 2006
  • The plastid transformation approach offers a number of unique advantages, including high-level transgene expression, multi-gene engineering, transgene containment, and a lack of gene silencing and position effects. The extension of plastid transformation technology to monocotyledonous cereal crops, including rice, bears great promise for the improvement of agronomic traits, and the efficient production of pharmaceutical or nutritional enhancement. Here, we report a promising step towards stable plastid transformation in rice. We produced fertile transplastomic rice plants and demonstrated transmission of the plastidexpressed green fluorescent protein (GFP) and aminoglycoside 3′-adenylyltransferase genes to the progeny of these plants. Transgenic chloroplasts were determined to have stably expressed the GFP, which was confirmed by both confocal microscopy and Western blot analyses. Although the produced rice plastid transformants were found to be heteroplastomic, and the transformation efficiency requires further improvement, this study has established a variety of parameters for the use of plastid transformation technology in cereal crops.